A clinically oriented computer model for radiofrequency ablation of hepatic tissue with internally cooled wet electrode.

PURPOSE: To improve the computer modelling of radiofrequency ablation (RFA) by internally cooled wet (ICW) electrodes with added clinically oriented features. METHODS: An improved RFA computer model by ICW electrode included: (1) a realistic spatial distribution of the infused saline, and (2) different domains to distinguish between healthy tissue, saline-infused tumour, and non-infused tumour, under the assumption that infused saline is retained within the tumour boundary. A realistic saline spatial distribution was obtained from an in vivo pig liver study. The computer results were analysed in terms of impedance evolution and coagulation zone (CZ) size, and were compared to the results of clinical trials conducted on 17 patients with the same ICW electrode. RESULTS: The new features added to the model provided computer results that matched well with the clinical results. No roll-offs occurred during the 4-min ablation. CZ transversal diameter (4.10 ± 0.19 cm) was similar to the computed diameter (4.16 cm). Including the tumour and saline infusion in the model involved (1) a reduction of the initial impedance by 10 - 20 Ω, (2) a delay in roll-off of 20 s and 70 - 100 s, respectively, and (3) 18 - 31% and 22 - 36% larger CZ size, respectively. The saline spatial distribution geometry was also seen to affect roll-off delay and CZ size. CONCLUSIONS: Using a three-compartment model and a realistic saline spatial distribution notably improves the match with the outcome of the clinical trials.

Multicenter Stratified Study From Spain About Acceptance of Living Liver Donation Among Veterinarian Students.

Veterinarians often hold decision-making positions in the public health care system, and therefore can influence public opinion about organ donation. The objective is to analyze the attitude of Spanish veterinarian students toward living liver donation (LLD) and to establish which factors have an influence on this attitude. METHODS: A sociological, interdisciplinary, multicenter, and observational study was carried out in the veterinarian students enrolled in Spain (n = 9000) (university academic year 2010-2011). A sample of 2854 students was stratified by geographic area and academic year. A validated questionnaire (PCID-DVH RIOS) was self-administered and completed anonymously. The questionnaire was applied to each academic year at compulsory sessions at randomly selected veterinary schools. Statistical analysis included t test, χ2 test, and logistic regression analysis. RESULTS: Completion rate was 94% (n = 2683); 89% (n = 2345) were in favor of related LLD, and 40% (n = 1053) supported unrelated LLD. The following variables were associated with a more favorable attitude: (1) age (P < .001), (2) sex (P < .001), (3) academic year (P < .001), (4) believing in the possibility of needing a transplant oneself in the future (P < .001), (5) attitude toward deceased donation (P < .001), (6) attitude toward living kidney donation (P < .001), (7) acceptance of a donated liver segment from a family member if one were needed (P < .001), (8) having discussed the subject with one's family (P = .009) and friends (P < .001), (9) a partner's opinion about the subject (P = .002), and (10) fear of the possible mutilation of the body after donation (P < .001). CONCLUSION: Spanish veterinary students have a favorable attitude toward LLD.

Veterinary Students' Attitude Toward Living Kidney Donation: A Multicenter Stratified Study From Spain.

Living kidney related donations (LKRD) should be promoted because of the current deficit of organs for transplantation. The objective of the study was to analyze the attitude of Spanish veterinary students toward LKRD, because they may influence public opinion in the future, and to determine the factors that condition it. METHODS: A sociological, interdisciplinary, multicenter, and observational study was carried out in the veterinary students enrolled in Spain (n = 9000) in the university academic year 2010-2011. A sample of 2815 students was stratified by geographical area and academic year. The students' attitude toward LKRD was assessed using a psychosocial validated questionnaire (PCID-DVR Rios), was self-administered, and was completed anonymously. Veterinary schools were randomly selected. The questionnaire was applied to each academic year at compulsory sessions. Statistical analysis was preformed using t test, χ2 test, and logistic regression analysis. RESULTS: The completion rate was 95% (n = 2683); 93% (n = 2504) were in favor of LKRD and 36% (n = 945) supported unrelated living kidney donation. The following variables were associated with having a more favorable attitude: (1) sex (P < .001), (2) being a student of southern universities (P = .03), (3) attitude toward deceased donation (P < .001), (4) having a father (P < .001) or a mother (P < .001) in favor of organ donation, and (5) having discussed the subject with friends (P = .03) or family (P = .02). However, only 60% would accept a kidney from a relative. CONCLUSION: Spanish veterinary students have a favorable attitude toward LKRD. However, only 60% would accept a kidney from a relative.

Attitude Toward Organ Donation and Transplantation in Veterinary Students: A Multicenter and Stratified Study in Spain.

BACKGROUND: Veterinarians often hold decision-making positions in the public health care system and can therefore influence public opinion about organ donation and transplantation (ODT). The aim of this work was to analyze the attitude of Spanish veterinary students toward ODT, because they may influence public opinion in the future, and to determine the factors that condition it. METHODS: This was a sociologic, interdisciplinary, multicenter, observational study in Spain. The study population was students studying for a veterinary degree in Spain (n = 9000), and a sample of 2815 students (confidence of 99% and precision of ±1%) was stratified by geographic area and academic year. A validated questionnaire of attitude toward ODT (PCID-DTO-Ríos) was self-administered and anonymous. RESULTS: Of the 2815 selected students (2790 plus the 0.9% per type of sample), 2650 completed the questionnaire (response rate, 94.14%): 83% (n = 2207) of the respondents were in favor of donation and 17% against. The following main variables were related to a favorable attitude: being female (odds ratio [OR], 0.752; P = .034); knowing a donor (OR, 1.834; P = .003); having discussed the matter with one's family (OR, 1.587; P = .002); having spoken about the subject in social circles with friends (OR, 1.633; P < .001), and being in favor of donating a deceased family member's organs (OR, 2.403; P < .001). CONCLUSIONS: Seventeen percent of Spanish veterinary students were not in favor of ODT. It is important to know the factors that determine their attitude, because this will make it possible to optimize the resources invested in campaigns to promote ODT and to take more specific action.

Long-term effectiveness of irreversible electroporation in a murine model of colorectal liver metastasis.

Irreversible electroporation (IRE) has recently gained in popularity as an ablative technique, however little is known about its oncological long-term outcomes. To determine the long-time survival of animals treated with a high dose of IRE and which histological changes it induces in tumoral tissue, IRE ablation was performed in forty-six athymic-nude mice with KM12C tumors implanted in the liver by applying electric current with different voltages (2000 V/cm, 1000 V/cm). The tumors were allowed to continue to grow until the animals reached the end-point criteria. Histology was harvested and the extent of tumor necrosis was semi-quantitatively assessed. IRE treatment with the 2000 V/cm protocol significantly prolonged median mouse survival from 74.3 ± 6.9 days in the sham group to 112.5 ± 15.2 days in the 2000 V/cm group. No differences were observed between the mean survival of the 1000 V/cm and the sham group (83.2 ± 16.4 days, p = 0.62). Histology revealed 63.05% ± 23.12 of tumor necrosis in animals of the 2000 V/cm group as compared to 17.50% ± 2.50 in the 1000 V/cm group and 25.6% ± 22.1 in the Sham group (p = 0.001). IRE prolonged the survival of animals treated with the highest electric field (2000 V/cm). The animals in this group showed significantly higher rate of tumoral necrosis.

Irreversible electroporation of the liver: is there a safe limit to the ablation volume?

Irreversible electroporation is a fast-growing liver ablation technique. Although safety has been well documented in small ablations, our aim is to assess its safety and feasibility when a large portion of liver is ablated. Eighty-seven mice were subjected to high voltage pulses directly delivered across parallel plate electrodes comprising around 40% of mouse liver. One group consisted in 55 athymic-nude, in which a tumor from the KM12C cell line was grown and the other thirty-two C57-Bl6 non-tumoral mice. Both groups were subsequently divided into subsets according to the delivered field strength (1000 V/cm, 2000 V/cm) and whether or not they received anti-hyperkalemia therapy. Early mortality (less than 24 hours post-IRE) in the 2000 V/cm group was observed and revealed considerably higher mean potassium levels. In contrast, the animals subjected to a 2000 V/cm field treated with the anti-hyperkalemia therapy had higher survival rates (OR = 0.1, 95%CI = 0.02-0.32, p < 0.001). Early mortality also depended on the electric field magnitude of the IRE protocol, as mice given 1000 V/cm survived longer than those given 2000 V/cm (OR = 4.7, 95%CI = 1.8-11.8, p = 0.001). Our findings suggest that ionic disturbances, mainly due to potassium alterations, should be warned and envisioned when large volume ablations are performed by IRE.

Cartilage resurfacing potential of PLGA scaffolds loaded with autologous cells from cartilage, fat, and bone marrow in an ovine model of osteochondral focal defect.

Current developments in tissue engineering strategies for articular cartilage regeneration focus on the design of supportive three-dimensional scaffolds and their use in combination with cells from different sources. The challenge of translating initial successes in small laboratory animals into the clinics involves pilot studies in large animal models, where safety and efficacy should be investigated during prolonged follow-up periods. Here we present, in a single study, the long-term (up to 1 year) effect of biocompatible porous scaffolds non-seeded and seeded with fresh ex vivo expanded autologous progenitor cells that were derived from three different cell sources [cartilage, fat and bone marrow (BM)] in order to evaluate their advantages as cartilage resurfacing agents. An ovine model of critical size osteochondral focal defect was used and the test items were implanted arthroscopically into the knees. Evidence of regeneration of hyaline quality tissue was observed at 6 and 12 months post-treatment with variable success depending on the cell source. Cartilage and BM-derived mesenchymal stromal cells (MSC), but not those derived from fat, resulted in the best quality of new cartilage, as judged qualitatively by magnetic resonance imaging and macroscopic assessment, and by histological quantitative scores. Given the limitations in sourcing cartilage tissue and the risk of donor site morbidity, BM emerges as a preferential source of MSC for novel cartilage resurfacing therapies of osteochondral defects using copolymeric poly-D,L-lactide-co-glycolide scaffolds.

Using wireless rumen sensors for evaluating the effects of diet and ambient temperature in nonlactating dairy goats.

Sixteen Murciano-Granadina dairy goats, provided with wireless rumen sensors for pH and temperature, were used to assess the rumen environment variations produced by extreme forage to concentrate diets (experiment 1) and climatic conditions (experiment 2). To avoid the interference of feed intake, goats were fed at maintenance level. Rumen sensors were inserted by surgery and programmed to collect and store rumen pH and temperature every 30min. In experiment 1, 8 dry goats (38.6±2.3kg of body weight) in tiestalls were divided into 2 groups and fed at maintenance level with 2 diets varying in forage-to-concentrate ratio [high forage (HF) 70:30; low forage (LF) 30:70] according to a crossover design. Diets were offered once daily for 4h and tap water (4 L, 9.8±0.4°C) was offered for only 30min at 6h after feeding. Rectal temperatures were recorded 3 times during the day. Rumen pH fell immediately after feeding, reaching a nadir depending on the diet (HF=6.35±0.07 at 11h after feeding; LF=6.07±0.07 at 6h after feeding) and being on average greater (0.31±0.06) in HF than LF goats. No diet effects were detected in rectal (38.2±0.1°C) and ruminal (38.9±0.1°C) mean temperatures, which were positively correlated. Rumen temperature dramatically changed by feeding (1.4±0.1°C) and drinking (-3.4±0.1°C), and 2h were necessary to return to the fasting value (38.2±0.1°C). In experiment 2, 8 dry goats (43.9±1.0kg of body weight) were kept in metabolic cages, fed a 50:50 diet and exposed to 2 climatic conditions following a crossover design. Conditions were thermoneutral (TN; 20 to 23°C day-night) and heat stress (HS; 12-h day at 37°C and 12-h night at 30°C). Humidity (40±5%) and photoperiod (light-dark, 12-12h) were similar. Goats were fed at maintenance level, the feed being offered once daily and water at ambient temperature was freely available. Intake, rectal temperature, and respiratory rate were recorded 3 times daily. Despite no differing in dry matter intake, rumen pH was lower in HS than in TN goats (-0.12±0.04). On the contrary, rumen temperature (0.3±0.1°C), rectal temperature (0.4±0.1°C), respiratory rate (77±5 breaths/min), and water intake (3.2±0.7 L/d) had a greater increase in HS than TN, which might indicate an altered microbial fermentation under high temperature conditions. In conclusion, wireless bolus sensors proved to be a useful tool to monitor rumen pH and temperature as affected by different feeding and climatic conditions.

Transitory improvement of articular cartilage characteristics after implantation of polylactide:polyglycolic acid (PLGA) scaffolds seeded with autologous mesenchymal stromal cells in a sheep model of critical-sized chondral defect.

Clinical translation of emerging technologies aiming at cartilage resurfacing is hindered by neither the appropriate scaffold design nor the optimal cell source having been defined. Here, critical-sized, chondral-only focal defects were created in sheep and treated with clinical-grade, co-polymeric poly-lactide:polyglycolic acid scaffolds either alone or seeded with 3.3 × 10(6) ± 0.4 × 10(6) autologous bone marrow-derived mesenchymal stromal cells and studied over 12 month follow-up. An untreated group was included for comparison. Second-look arthroscopy performed at 4 months post-treatment evidenced the generation of neocartilage of better quality in those defects treated with cells. However, macroscopic scores in the cell-treated group declined significantly from 7.5 ± 2.3 at 4 months to 3.1 ± 2.6 (p = 0.0098) at 12 months post-treatment, whereas the other two experimental groups remained unaltered during 4-12 month post-treatment. The effectiveness of the cell-based approach proposed in this study is thus restricted to between months 1 and 4 post-treatment.

Effects of an intravenous bolus of alfaxalone versus propofol on intraocular pressure in sheep.

The objective of this prospective study was to determine the effects of a single intravenous bolus of alfaxalone in 2-hydroxypropyl-ß-cyclodextrin and propofol on the intraocular pressure (IOP) in sheep. Ten Ripollesa sheep with a bodyweight of 48.5 (6.8) kg (mean [sd]) were used in the study. Twenty-four hours before the experimental procedure, a complete ophthalmic examination was performed in all animals. The day of the study, intravenous alfaxalone (2 mg/kg) or propofol (6 mg/kg) was randomly administered in a cross-over design, with a washout period of two weeks. Measurements of IOP, globe position and pupil size were obtained at basal time, before induction (time 0) and at two, five, 10, 15, 20, 30, 45, 60, 90 and 120 minutes after drug administration. Occasional side effects and time to standing were also noted. Intravenous administration of alfaxalone and propofol in sheep resulted in no alteration of IOP. Nevertheless, a decrease in the pupil size was observed in both groups. This present study shows that alfaxalone and propofol, administrated as a single intravenous bolus, are good options for maintaining IOP during anaesthesia in sheep, although marked miosis was observed after administration.

Prepubertal goat oocytes from large follicles result in similar blastocyst production and embryo ploidy than those from adult goats.

Developmental competence of oocytes from prepubertal females is lower than those from adult females. Oocyte development competence is positively related to follicular diameter. Most of the follicles of prepubertal goat ovaries are smaller than 3 mm. The aim of this study was to compare oocytes of two follicle sizes (< 3 mm and ≥ 3 mm) from prepubertal goats with oocytes from adult goats in relation to their in vitro production and quality of blastocysts. Oocytes from prepubertal goats were obtained from slaughterhouse ovaries and selected according to the follicle diameter whereas oocytes from adult goats were recovered in vivo by LOPU technique without prior selection of follicle size. COCs were IVM for 27 h, IVF at the conventional conditions with fresh semen and presumptive zygotes were cultured in SOF medium for 8 days. Blastocysts obtained were vitrified and after warming their blastocoele re-expansion and the ploidy by FISH technique were assessed. We found significant differences between blastocysts yield of oocytes recovered from follicles smaller than 3 mm of prepubertal goats compared to those from adult goats (5.45% vs 20. 83%, respectively) however, these differences disappear if oocytes were recovered form large follicles (18.07%). A total of 28 blastocysts were analysed and 96.43% showed mixoploidy. Age did not affect the number of embryos with abnormal ploidy or blastocyst re-expansion after warming. Furthermore, the percentage of diploid blastomeres per embryo was similar in the 3 groups studied, adult, prepubertal from follicles ≥ 3 mm and < 3 mm (68.6%, 80.8% and 73.6%, respectively). In conclusion, IVP of blastocysts coming from follicles larger than 3 mm of goats 45 days old were not different to the blastocysts produced from adult goats, both in terms of quantity and quality.

Plasma buprenorphine concentrations after the application of a 70 microg/h transdermal patch in dogs. Preliminary report.

The objective of the present study was to evaluate the plasma concentrations and pharmacokinetics of buprenorphine after transdermal application in dogs (n = 4). A 70 microg/h transdermal buprenorphine patch was applied to the ventral abdomen of four healthy beagles. Blood samples were collected through a preplaced jugular catheter before and at 1, 2, 4, 8, 12, 24, 36, 48 and every 6 h until 108 h after the patch application. Plasma buprenorphine concentrations were measured using a (125)I-labelled radioimmunoassay (RIA) assay. No adverse effects were observed in any of the dogs. Concentrations of buprenorphine were detected in plasma after the application of the transdermal buprenorphine patch on the four experimental animals. Buprenorphine plasma concentrations increased during the first 36 h and then remained in the 0.7-1.0 ng/mL range during the study period. A decrease in plasma buprenorphine concentration was not observed during the study. Although analgesia could not be demonstrated the present study shows the ability of buprenorphine transdermal delivery systems developed for human use to deliver measurable concetrations of buprenorphine in dogs.

Evaluation of a dynamic in vitro model to simulate the porcine ileal digestion of diets differing in carbohydrate composition.

The aim of the study was to assess the ability of a dynamic in vitro model to determine the digestibility of OM, CP, and starch compared with a validated, static, in vitro method and in vivo ileal digestibility obtained from growing pigs fitted with a T-cannula. Five experimental diets with different carbohydrate types and level were assessed: a standard corn-based diet (ST) or the same diet with coarse ground corn (CC), 8% sugar beet pulp (BP), 10% wheat bran (WB), or 8% sugar beet pulp and 10% wheat bran (HF). In the in vivo experiment, diets CC and HF reduced (P = 0.015) ileal digestibility of OM compared with the ST diet. The inclusion of sugar beet pulp reduced (P = 0.049) ileal CP digestibility of the BP diet. This reduction was not statistically significant when sugar beet pulp was combined with the wheat bran in the HF diet. No differences were shown for in vivo starch digestibility among diets. With the static in vitro method, the OM disappearance was greater than that observed in the in vivo experiment. In this static method, the BP and HF diets reduced (P = 0.004 and < 0.001, respectively) the disappearance of the OM compared with the ST diet. The coarse grinding of corn did not alter OM digestibility but decreased (P = 0.005) the starch digestibility. The R(2) between the in vivo results and the static in vitro methods for OM and starch digestibility was 0.99 when the CC diet was not considered. The dynamic in vitro model yielded OM and CP digestibility coefficients comparable with those obtained in vivo for the ST and CC diets. However, the values were considerably affected by the incorporation of the fibrous ingredients. Diets BP, WB, and HF had decreased (P = 0.009, 0.058, and 0.004, respectively) OM digestibility compared with the ST diet. Protein digestibility was also decreased (P < 0.001, P = 0.019, and P = 0.003, respectively) with the BP, WB, and HF diets compared with the ST diet. However, digestibility was decreased to a greater extent in the BP diet than in the WB and HF diets, both of which contained wheat bran. The R(2) between the dynamic in vitro model and the in vivo results for CP digestibility was 0.99 when the CC diet was not considered. No differences were detected for starch digestibility among the diets with the dynamic in vitro model. This dynamic in vitro model yielded ileal digestibility results comparable with those obtained in vivo for CP and OM with a corn-soybean diet, or with a diet including coarse corn, but it underestimated digestibility when fibrous ingredients were included in the diet.

Microsomal prostaglandin E synthase-1, which is not coupled to a particular cyclooxygenase isoenzyme, is essential for prostaglandin E(2) biosynthesis in vascular smooth muscle cells.

BACKGROUND: Prostaglandin (PG) E(2) induces expression of matrix metalloproteinases and angiogenic factors, thereby contributing to plaque instability. OBJECTIVE: To study the influence of cyclooxygenase (COX) and PGE synthase (PGES) isoenzyme expression on PGE(2) and PGI(2) biosynthesis in vascular smooth muscle cells (VSMC) in culture. METHODS: Cells were treated with human recombinant IL-1beta over different periods of time. Expression of PGI synthase, and COX and PGES isoenzymes was determined by real-time reverse transcriptase polymerase chain reaction and immunoblotting. Biosynthesis of prostanoids from exogenous or endogenous substrate was analyzed by high-performance liquid chromatography or enzyme-immunoassay after incubation of cells with labeled arachidonic acid or thrombin, respectively. RESULTS: Cytosolic PGES and microsomal PGES (mPGES) -1 and -2 were expressed in VSMC. PGES activity was mainly linked to mPGES-1. IL-1beta induced COX-2 and mPGES-1 with a different time course. VSMC ability to synthesize PGE(2) and PGI(2) fitted mPGES-1 and COX-2 expression, respectively. The ability of VSMC to produce PGI(2) was downregulated by mPGES-1 expression and was restored when mPGES-1 expression was silenced. Results from COX-1 and COX-2 silencing and selective inhibition showed that both COX-1 and COX-2 were involved in the biosynthesis of PGE(2) and their relative contribution depended on the time of incubation with IL-1beta. CONCLUSIONS: mPGES-1 is the main PGES responsible for PGE(2) biosynthesis by VSMC and its induction downregulates VSMC ability to produce PGI(2.) These results support the concept that under inflammatory conditions VSMC could significantly contribute to plaque instability and that mPGES-1 may be a target for therapeutic intervention in patients with cardiovascular risk.

Extremity amniotic band syndrome in fetal lamb. I: An experimental model of limb amputation.

OBJECTIVE: The objective of the study was to reproduce severe forms of extremity amniotic bands, which result in amputation or severe lesions. STUDY DESIGN: Right limbs of 5 60-day gestational age pregnant sheep were ligated with silk suture at the infracondylar level. Left limbs were used for paired comparison. The limbs obtained from term fetuses were analyzed morphologically, radiologically, and histologically. RESULTS: The ligated limbs showed an amputation or quasiamputation. Four extremities showed a necrotic pattern and 4 an edema pattern. CONCLUSION: A model of severe extremity amniotic bands that reproduces the effect of the extremity amniotic bands in the human fetus was developed. Future applications of this new model could be an experimental study of in utero salvage of limbs affected with severe extremity amniotic bands in the ovine fetus.

Inflammatory biomarkers of coronary atheromatous plaque vulnerability.

In the last decade, compelling evidence has evolved at both the basic science and clinical level for the implication of inflammation in the pathogenesis of atherosclerosis and its complications. The composition of the atherosclerotic plaque, rather than the degree of stenosis, is now recognized as a pivotal feature in determining plaque vulnerability and hence the risk of acute coronary ischaemic events. Current evidence supports a key role for inflammation in all phases of the atherosclerotic process, from plaque formation through to progression and, ultimately, the thrombotic complications of atherosclerosis. The growing appreciation of the role of inflammation in atherogenesis has focused attention on whether circulating levels of inflammatory biomarkers may help to identify those at risk of future cardiovascular events. In addition, the protective effects of a variety of interventions, such as statins, aspirin, and fibrates, are often associated with the evidence of reduced inflammation, further strengthening the notion that inflammation and the acute complications of atherosclerosis are causally related. The present review describes the pathophysiology of atheromatous plaque vulnerability and discusses the clinical use of inflammatory biomarkers for prognostic stratification of patients with acute coronary syndromes.

Nuevos conceptos sobre la enfermedad renal crónica y sus implicaciones / No disponible

La National Kidney Foundation americana ha definido recientemente criterios que definen un nuevo concepto denominado enfermedad renal crónica. Más o menos discutibles, suponen un importante método de estandarización de rápida aceptación internacional. En este artículo se revisa la nueva definición y estadios, los métodos de diagnóstico y evaluación, sus complicaciones y asociaciones, así como sus factores de progresión a estadios terminales con necesidad de tratamiento sustitutivo. Asimismo se establece el importante nuevo vínculo de la enfermedad renal crónica como factor de riesgo cardiovascular (AU)

The American National Kidney Foundation has recently defined new criteria to define the concept of chronic Kidney disease (CKD). Althougt they are partially under discussion, they have become a very helpful way of standarization and widely internationally accepted. In this article, we review the new definition and stages, the diagnostic and evaluation methods, complications and associations, as well as progression factors to end stage renal disease. It is also underlined the important newly recognized link between chronic kidney disease and cardiovascular risk (AU)